The overall purpose of this NIH Shared Instrument Grant application is to provide unique and crucial instrumentation to facilitate and enhance the research efforts of 8 Principal Investigators on 14 independent NIH research projects at the Mount Sinai School of Medicine. The specific request is for an Automated DNA Sequencing Facility consisting of an ABI Model 373A Automated DNA Sequencer with printer and quantitation/genotyping software to generate, collect and analyze data, a Beckman Biomek 2000 BioRobotics Workstation to automate sequencing reaction preparation, and a Sun SPARCsystem with tape backup which is required for sequence editing/proofreading, data distribution and data storage. The SPARCsystem computer will be on our Institutional Ethernet backbone and can support multitasking for remote access to finished sequence data and the Staden DNA sequence manipulation programs. The Sequencing Facility will utilize Taq cycle sequencing which provides ease of use with the widest variety of templates. The dye-terminator chemistry of the Taq cycle sequencing protocol permits users to employ any primer they have in their stockpile. Due to the varied research needs of the large group of Major Users, our present Operating Plan provides for both user and core template preparation. The increased flexibility of this approach should outweigh the sequencing consistency gained by locking users into specific vector systems and/or primers. The usefulness of this facility is enhanced by the fact that the Department of Human Genetics also runs an Oligonucleotide Synthesis core facility and can thus provide a rapid turnaround time for generating primers for sequence walking and other needs. The proposed Sequencing Facility would be administered, operated and maintained in the Department of Human Genetics as a sequencing and genotyping core facility to primarily support the heavy sequencing needs of established and young Departmental Faculty. A cost- recovery charge will be assessed only for supply expenditures. By automating the analysis of over 6,000 sequencing reactions per year, we expect an enhanced efficiency in the conduct of a large number of research projects and a reduction in errors of pipetting and manipulation due to the reproducibility and accuracy of these automated methods. Excellent hardware, software and network connectivity support is also available through the Academic Computing Division at Mount Sinai which will facilitate optimal use of our system and provide access to Internet services. To support user's needs for data analysis, the Department of Human Genetics also administers the GCG package and other software such as phylogenetic analysis programs and EMaiI servers that run on our networked institutional VAX computers. In summary, the requested sequencing instrumentation will fit synergistically with our existing resources to provide state-of-the-art genomics support for NIH-funded research at Mount Sinai.